An Experimental Bacteriostasis of the Banxiaxiexin Decoction and 7 Kinds of Single Taste Traditional Chinese Medicine on Helicobacter Pylori ...
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American Journal of Internal Medicine
2020; 8(3): 138-142
http://www.sciencepublishinggroup.com/j/ajim
doi: 10.11648/j.ajim.20200803.18
ISSN: 2330-4316 (Print); ISSN: 2330-4324 (Online)
An Experimental Bacteriostasis of the Banxiaxiexin
Decoction and 7 Kinds of Single Taste Traditional Chinese
Medicine on Helicobacter Pylori Resistant Strains in Vitro
Zhiwei Qu1, *, Mingjun Yu2, Airong Zhang2, Ailin Wang3, Liyuan Shun3
1
Department of Traditional Chinese Medicine, The Affiliated Hospital of Beihua University, Jilin, China
2
Department of Gastroenterology, The Affiliated Hospital of Beihua University, Jilin, China
3
Department of Laboratory Medicine, Beihua University, Jilin, China
Email address:
*
Corresponding author
To cite this article:
Zhiwei Qu, Mingjun Yu, Airong Zhang, Ailin Wang, Liyuan Shun. An Experimental Bacteriostasis of the Banxiaxiexin Decoction and 7 Kinds
of Single Taste Traditional Chinese Medicine on Helicobacter Pylori Resistant Strains in Vitro. American Journal of Internal Medicine.
Vol. 8, No. 3, 2020, pp. 138-142. doi: 10.11648/j.ajim.20200803.18
Received: April 19, 2020; Accepted: May 3, 2020; Published: May 28, 2020
Abstract: Objectives: To research experimental bacteriostasis on the Banxiaxiexin decoction and 7 kinds of single taste
Traditional Chinese Medicine (TCM) for Helicobacter pylori resistant strains in vitro. Methods: Collecting specimens of gastric
mucosa in patients with upper gastrointestinal disease, separated and identified 8 strains of Hp resistant strains, the E-test
methods of metronidazole and clarithromycin, drug resistance status of amoxicillin, selected eight strains of clinical drug
resistance strains; Using liquid dilution method to determine the Banxiaxiexin decoction and 7 kinds of single Chinese medicine
clinical drug-resistant strains of 8 strains of antibacterial effect in vitro. Results: Banxiaxiexin decoction MIC is 0.31 mg/ml,
MIC of Radix scutellariae is 0.09 mg/ml, Rhizoma coptidis MIC 0. 16 mg/ml, Glycyrrhizin realness MIC 0. 31 mg/ml, Ginseng
MIC 0. 75 mg/ml, there are four of seven kinds of single taste Traditional Chinese Medicine of TCM has bacteriostatic action, the
order of antibacterial strength: Scutellaria baicalensis>Coptis chinesis>Glycyrrhizin realness >Ginseng, three kinds of Pinellia,
Jujube and dry Zingibe in vitro for H. P ylori no inhibition, the method of MIC with liquid dilution in this decoction were
determined, the decoction of resistant isolates average MIC isolates was 0.20±0.09mg/ml, the results showed that this decoction
have stronger antibacterial effect on eight kinds of drug resistant strains in vitro. Banxiaxiexin decoction and 4kinds of single
taste TCM have stronger antibacterial effect on drug resistant strains in vitro. Conclusion: Banxiaxiexin decoction and 4 kinds of
single TCM have strong bacteriostatic effect on the bacteria resistance of Helicobacter pylori strains in vitro, therefore, we
suggest that this decoction could be used as an effective method in the treatment of resistant HP associated gastritis.
Keywords: Helicobacter Pylori, Drug Resistance, Banxiaxiexin Decoction, Bacteriostatic Effect
Shang-han lun of prescriptions for Pinellia, Scutellaria
1. Introduction baicalensis, Zingiber, Ginseng, Glycyrrhiza uralensis, Coptis
Helicobacter pylori is the cause of chronic active gastritis chinesis, Jujube, the function may be modulated the cold and
and duodenal ulcer. It is given priority to antibiotic heat balance, eliminated swelling of the liver disease and
combination scheme of Hp infection with effective treatment dissipated nodules, Indicating on cold and heat mixed in the
of western medicine [1], but along with the increase in ruffian Syndrome as well as abdomen with swelling of the
antibiotic use, side effects and the resistant strains are also liver disease, but abdominal fullness without pain, or vomiting,
more and more [2-3]. From the perspective of Traditional bowel diarrhea, and yellowish greasy tongue. Clinical
Chinese Medicine to study the prevention and control drug commonly we have used in Weak qi, cold and heat mutual
resistant strains has opened up a new way, Banxiaxiexin knot, such as early acute or chronic gastroenteritis, chronic
decoction from zhang zhongjing in Chinese han dynasty [4], colitis, chronic hepatitis, cirrhosis of the liver [5-7]. We have
American Journal of Internal Medicine 2020; 8(3): 138-142 139
curative effects on treating Helicobacter pylori related gastric gastroscope, a piece of antrum and body of stomach were
disease in clinical observation of the decoction, especially taken respectively with biopsy forceps within 5cm from the
resistant Helicobacter pylori cases, but the specific front of pylorus, used sterile forceps to put the specimen into a
mechanism is unclear, therefore, we have researched centrifuge tube filled with delivery solution, placed it in 4°C
experimental bacteriostasis on the decoction and 7 kinds of for preservation and transferred to the laboratory. In the super
single taste Traditional Chinese Medicine (TCM) for clean table of aseptic room, a piece of gastric mucosa tissue
Helicobacter pylori resistant strains in vitro, the purpose is to were taken, pressed the mucosa tissue with inoculation ring
explore the decoction and 7 kinds of single Chinese medicine and smeared it on skirrow culture medium (Shanghai Yubo
resistance mechanism of H. pylori resistant strains, could be Biological Technology Co., Ltd, China); poured skirrow
provided the objective basis for the prevention and treatment culture medium into anaerobic tank for 3 days, HP culture
of H. pylori resistant strains. conditions: 37°C, 10% CO2, 5% O2, 85% N2. After 3-5 days
of culture, the transparent needle like colonies on the agar
2. Methods plate were selected, transferred to a new skirrow selective
medium, and the strains were purified and enriched.
2.1. Patients and Sample Preparation
2.4. Identification of HP Bacteria
A total of 105 patients seen at the Department of
Gastroenterology, Affiliated Hospital of Beihua University, Morphological examination: the HP colonies were
China, from July 2016 to August 2019 were included. The inoculated by scribing line showed transparent needle tip
following cases were excluded: gastrointestinal cancer, shape, neat edge, central protuberance and diameter of 1. 2
surgical history, pregnancy and lactation, and so those with ram colony; a translucent layer of lichen when inoculated with
severe disease. Before gastroscopy, all the patients were large amount of bacteria. Select the typical HP colony,
confirmed that had not treated by any Hp eradication therapy smeared it in a drop of normal saline, dry it, and then gram
(antibiotics, acid-suppressing drugs, NSAIDs or bismuth) stain it, and observed it under ordinary microscope. Seeing
within one month. The antibacterial test was performed with gram-negative S-shaped Campylobacter or Brevibacterium
disc agar diffusion (K-B method). Drug resistant strains I ~ (see Figure 1). It can be preliminarily determined as HP. 2)
VII were isolated and cultured from 105 patients' gastric Biochemical examination: taking HP colony for urease,
mucosa, resistant to metronidazole, amoxicillin, catalase and oxidase test (Nanjing dulai Biotechnology Co.,
clarithromycin, ornidazole, levofloxacin, moxifloxacin and Ltd, China). If all three biochemical reactions are positive, HP
rifampin, as well as Susceptibility testing to etronidazole were determined as:
amoxicillin, clarithromycin, ornidazole, levofloxacin,
moxifloxacin and rifabutin was performed with disc agar
diffusion method (Bio Merieux, France), results determine
reference [8-9]. Strains were classified as resistant when the
MIC (minimal inhibitory concentration) was ≥8 µg / ml
(etronidazole); ≥0.5µg / ml (amoxicillin); ≥1µg / ml
(clarithromycin); ≥2µg / ml (ornidazole); ≥2µg / ml
(levofloxacin); ≥2µg / ml (moxifloxacin); ≥2µg / ml
(rifampicin). The standard strain NCTC11637 was donated by
Professor Hu Fulian of the First Affiliated Hospital of Peking
University.
2.2. Chinese Medicine Reagent
We have selected the Chinese medicinal materials of
banxiaxiexin decoction as follows: Put Pinellia (9 g),
Scutellaria baicalensis (6 g), Zingiber (6 g), Ginseng (6 g), Figure 1. Morphology of Helicobacter pylori 10x100.
Glycyrrhiza uralensis (6 g), Coptis chinensis (3 g), Jujube (3 g)
in 500ml water, simmered for 2 h, filtered residue and added HP preservation: We have selected the third generation HP
water to continue boiling for 30 min, filtration to get the final of enrichment culture, and preserved it in its logarithmic
concentration (each 100ml liquid contained 10 g of original growth period (General 48 hours); added 3-5ml of
drugs), Chinese medicine, Pinellia, Radix scutellariae, preservation solution (30% glycerol bouillon) to elute the
Ginseng, Rhizoma coptidis, Radix glycyrrhizae, Jujube, Dried bacteria on the agar surface, used aseptic straw to separate the
ginger, were purchased from Sanjiu Pharmaceutical Co., Ltd, bacterial solution into 1.5ml aseptic eppend of tube or
China. cryopreservation tube, and quickly placed the strain in - 80°C
refrigerator for preservation.
2.3. Isolation and Culture of H. Pylori
HP recovery: the bacteria were taken out directly from the
Specimen collection and HP separation and culture: through refrigerator, unfreezed at room temperature, used the sterile
140 Zhiwei Qu et al.: An Experimental Bacteriostasis of the Banxiaxiexin Decoction and 7 Kinds of Single Taste
Traditional Chinese Medicine on Helicobacter Pylori Resistant Strains in Vitro
elbow pipette to absorb the bacteria solution, dropped it on the bacterial suspension was added, and the eighth hole was
skirrow selective culture medium, used the L-shaped glass rod negative control. Microaerobic environment, 120 R / min,
to evenly coat the bacteria solution, and putted the culture dish shaking culture 48 h. The turbidity of the culture plate was
into the incubator for incubation. the conditions were the same observed with naked eyes, and the samples were stained and
as before. Drug sensitivity test: we have collected fresh HP biochemical identified. At the same time, 100 µ l was taken to
from pure culture for 48-72 hours and fully grind it into sterile the solid medium for further culture for 72 hours, and the
saline, adjust the concentration to 2 × 108cfu / ml, dipped the growth of the colony was observed. If there was no bacterial
sterile cotton swab in the bacterial solution, evenly coated the growth, it was judged that it had bacteriostatic effect, If it is
whole agar surface, stand, slightly dry, took the E-test strip found that bacteria grow on the medium containing the highest
containing antibiotics and laid it flat on the agar surface (1 / 90 concentration of 0.31mg/ml, its MIC value is calculated as
mm plate), 37°C, and cultured in microaerobic environment 0.31mg/ml.
for 72 hours, the results showed that the minimum inhibitory
concentration (MIC) was the scale at the junction of the 3. Results
ellipsoidal bacteriostatic ring and the test strip.
The results of Banxiaxiexin decoction and 7 kinds of single
2.5. Determination of MIC taste TCM for HP standard strains in vitro bacteriostasis
MIC were determined by liquid multiple dilution method: experiment MIC are shown in table 1 the decoction and 7
experiment study on antimicrobial Banxiaxiexin decoction kinds of single Chinese medicine MIC were determined by
and seven kinds of single herbs resistant strains of H. pylori in liquid dilution, the results showed that the Banxiaxiexin
vitro assays were proformed. Concentration Banxiaxiexin decoction and 7 kinds of single taste TCM for HP, strong
decoction were 10mg/ml, the concentration of each herb was: bacteriostatic activity of Banxiaxiexin decoction MIC is 0.31
Pinellia 15mg/ml, Scutellaria baicalensis 6mg/ml, Zingiber mg/ml, Radix scutellariae MIC 0.09 mg/ml, Rhizoma coptidis
12mg /ml, Ginseng12 mg/ml, Glycyrrhiza uralensis 10mg/ml, MIC 0. 16 mg/ml, Licorice MIC 0. 31 mg/ml, Ginseng MIC 0.
Coptis chinensis 10 mg/ml, Jujube 6 mg/ml. Took 8 holes of 75 mg/ml, there are four kinds of Chinese medicine in 7 kinds
48 holes microtiter plate, added 1 ml of liquid medium into of single taste TCM, the strength of inhibitory effects
each hole, and 200 µ l of the above preparation solution into followed as: the Scutellaria baicalensis, Coptis chinensis,
the first hole, shaking well, and diluted to the eighth hole by Glycyrrhiza uralensis, Ginseng, However, the strongest one is
multiple ratio. the drug concentration were 128, 64, 32, 16, 8, Scutellaria and weakest Ginseng, but no inhibition in three
4, 2 mg / ml in turn. In each tube, 10 µ l of liquid culture HP kinds of Pinellia, Jujube and Dry zingibe in vitro.
Table 1. MIC value of Banxia Xiexin Decoction and 7 kinds of single Chinese herbs against H. pylori (mg/ml).
Chinese pinyin English name Latin name MIC limits MIC50
Banxia Xiexin Tang Banxia Xiexin Decoction Banxia Xiexin Decoction 32~128 0.31
Huang qin Baikal Skullcap Scutellaria baicalensis 64~128 0.09
Huang lian Coptis chinensis Coptis chinensis Franch 64~128 0. 16
Gan cao Licorice Glycyrrhiza uralensis 32~128 0. 31
Ren shen Ginseng Panax schinseng 16~128 0.75
Banxia Pinelliae Rhizoma Pinellia ternatab 2~128 0
Aan jiang Dried ginger Zingiber officinale Rosc 2~128 0
Da zao Jujube Ziziphus jujuba Mill 2~128 0
strains in bacteriostatic activity, standard strains of MIC is
Table 2. MIC of Banxia Xiexin Decoction to 8 drug resistant strains by 0.31 mg/ml, Isolates 1 MIC 0.31 mg/ml, Isolates 2 MIC 0. 15
continuous dilution of solid medium.
mg/ml, Isolates 3 MIC 0. 08 / mg/ml, Isolates 4 MIC 0. 15
Bacterial strain MIC limits MIC50 mg/ml, Isolates 5 MIC 0. 31 mg/ml, Isolates 6 MIC 0. 15
Standard strain 32~128 0.31 mg/ml, Isolates 7 MIC 0. 31 mg/ml, Isolates 8 MIC 0.15
Isolate 1 32~128 0.31
mg/ml, the average MIC were 0.20±0.09mg/ml (x±SD), So
Isolate 2 16~128 0. 15
Isolate 3 8~128 0. 08 that the decoction have strong effects on antibacterial in vitro
Isolate 4 16~128 0.15 against 8 kinds of drug resistant strains.
Isolate 5 32~128 0.31
Isolate 6 16~128 0.15
Isolate 7 32~128 0. 31 4. Discussion
Isolate8 16~128 0. 15
At present western medicine for Helicobacter pylori
This decoction to eight kinds of resistant strains of eradication therapy is most typically applied to clinical
bacteriostatic effect in vitro bacteriostasis experiment MIC is antibiotic therapy, Containing the PPI triple therapy is one of
shown in table 2, the MIC of Banxiaxiexin decoction were the most admired the preferred therapy (omeprazole +
determined by liquid dilution method, the results showed that metronidazole, amoxicillin and clarithromycin in any [10-11].
Banxiaxiexin decoction is stronger to 8 kinds of resistant Although there are a lot of western medicine antibioticAmerican Journal of Internal Medicine 2020; 8(3): 138-142 141
therapy clinical studies have shown that its eradication of Acknowledgements
Helicobacter pylori is very effective, but there are still many
problems of therapy, including poor patient compliance, drug Professor Mingcheng Li from College of Medical
side effects, may be caused disorder in patients with Laboratory Science of Beihua University have contributed to
gastrointestinal flora, and broke out Superinfection the revision and finalization of later manuscripts, and
repeatedly, especially because of the rapid increase of Professor Haiying Gu from department of microbiology of
drug-resistant strains in recent years, the clinical efficacy of Hainan medical college have given great help to
antibiotic therapy has declined in varying degrees [12]. For microbiological test.
these reasons, many researchers at home and abroad are
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